Communication

Encapsulation and perfusion of mitochondria in agarose beads for functional studies with 31P-NMR spectroscopy

  1. Nicolai M. Doliba1,
  2. Suzanne L. Wehrli2,
  3. Andriy M. Babsky1,
  4. Nataliya M. Doliba1,
  5. Mary D. Osbakken M.D, Ph.D.3,*

Article first published online: 12 DEC 2005

DOI: 10.1002/mrm.1910390502

Issue

Magnetic Resonance in Medicine

Magnetic Resonance in Medicine

Volume 39, Issue 5, pages 679–684, May 1998

Additional Information

How to Cite

Doliba, N. M., Wehrli, S. L., Babsky, A. M., Doliba, N. M. and Osbakken, M. D. (1998), Encapsulation and perfusion of mitochondria in agarose beads for functional studies with 31P-NMR spectroscopy. Magnetic Resonance in Medicine, 39: 679–684. doi: 10.1002/mrm.1910390502

Author Information

  1. 1

    University of Pennsylvania, Children's Hospital of Philadelphia, Philadelphia, Pennsylvania

  2. 2

    Division of Metabolism, Children's Hospital of Philadelphia, Philadelphia, Pennsylvania

  3. 3

    Pharmaceutical Research Institute, Bristol-Myers Squibb, Princeton, New Jersey

*Bristol-Myers Squibb Pharmaceutical Research Institute, PO Box 4000, Room J3225, Princeton, NJ 08543-4000

Publication History

  1. Issue published online: 12 DEC 2005
  2. Article first published online: 12 DEC 2005
  3. Manuscript Accepted: 8 JAN 1998
  4. Manuscript Revised: 6 JAN 1998
  5. Manuscript Received: 23 JUL 1997

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Keywords:

  • mitochondria;
  • ATP production;
  • 31P NMR

Abstract

An NMR method to study on-line mitochondrial function was developed. Mitochondria were maintained in a stable physiologic state in agarose beads that were continuously superfused with oxygenated buffer at 28°C. Oxidative function of both heart and liver mitochondria was evaluated with 31P NMR at 9.4 T using pyruvate plus malate as substrate. This method allows clear resolution of adenosine triphosphate-γ (ATPγ) and adenosine diphosphate-β (ADPβ) phosphate signals, whereas a signals of ATP and ADP overlap. ATP production by mitochondria was documented to be very sensitive to different interventions (hypoxia, ischemia, carbonyl cyanide p-(tri-fluoromethoxy)phenylhydrazone (FCCP)) and depended on the ADP concentration in supervision medium. These data demonstrate that the new application of NMR to study mitochondrial function can discriminate, on-line, between several physiologic and biochemical processes in intact physiologically stable mitochondria.

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